Study On The Broad-spectrum Detection Of Quinolone Drugs

Quinolones drugs, because of its chemical structure characteristics, all havesuperb stability, can tolerate high temperature and high humidity environment, canalso preserve at room temperature for a long time. Therefore, if quinolones drugs havehigh residual in animal food, they will enter the human body, cause gastrointestinaland central nervous system adverse reactions、Toxic action for the heart、skinreactions、Phototoxicity and Cartilage toxicity and so on. In recent years, the residuesof quinolones drugs in animal tissue have caused extensive attention.Through norfloxacin and N-（4-Bromobutyl） phthalimide’s friedel FriedelCraftssynthetize the norfloxacin derivatives, this synthetic method have no reports in theexisting literature, is very simple and also has a high production rate. The difficult ofthis method is how to control the reaction temperature and time, and the question ofreaction products’ purification. Then norfloxacin derivatives couple with the BSA andOVA by Glutaraldehyde method, synthetize immunogen and coating antigen. AfterUV identification and the ELISA experiment, we are sure about the hapten and theprotein couple success. Now we immune nine Kunming mice respectively to preparepolyclonal antibody, after screening, measure cross response rate roughly to chooseantiserum which has higher titer and better inhibition, then conduct the follow-upELISA experiment.Through the use of serum as the foundation, base on polyclonal antibody ofindirect ELISA method preliminarily. By optimizing coat liquid、antibody diluent、confining liquid、 antibody reaction time、 the volume ratio of antibodyandstandardsolution etc, finally we get the ideal standard curve, achieve betterexperiment results. Ultimately, detection limit is0.0125ng/mL、the half inhibition(IC₅₀)is0.30515ng/mL, correlation coefficient of the standard curve is0.9923.Detection limit of the negative samples such as pig kidney、chicken、chicken liver is0.9986ng/mL、0.4928ng/mL、0.6492ng/mL.The paper also from sensitivity、specificity、accuracy and precision experimentalmethod to appraise ELISA method. The recovery contains63%⁹5%, the variation coefficient is less than20%. In addition to the cross reactive rate with Orbifloxacinlower than5%, and the rest of the another ten kinds of quinolones drugs are between7.21%⁹1.23%, accord with multi residue detect demand of ELISA, it also furtherprove the modification process about nofloxacin is successful, achieve the expectedeffect, meet the broad spectrum detection demand of quinolones drugs.